Objective To amplify and sequence the distinctive norB gene of Nitrobacterium. Methods The norB gene was amplified by PCR and was cloned into T-vector and then transformed into E.coli JM109. After white-and-blue selection, positive colonies were sequenced with T7 sequencing primers by Takara Company. Spliced with Vector NTI Suite software, the homologous analysis of sequences were conducted in Genbank through Internet. Results The results showed that all of norB genes obtained from the nitrite-oxidizing bacteria were homologous with the norB gene of Nitrobacterium hamburgensis in Genbank. The homology ranged from 96% to 98%, and there were no frameshift mutations, which guaranteed the correct ORF. Conclusion The obtained genes are norB genes indeed.

Objective To screen and seperate nitrobacteria that could be used to treat nitrogenous compound pollution.Methods Autotrophic nitrite-oxidizing microorganisms were screened by enrichment culture from soil.After morphological and biological detection,primers were designed to detect the distinctive gene norB of nitrobacteria.The PCR production was se-quenced,and the homology was identified according to the sequence in Genbank.Results One strain of bacterium was screened that could oxidize nitrite.This bacterium was small,light-yellow bacilli.The expected-size DNA fragments could be amplicated by PCR.The sequence of a PCR production was blasted in Genbank and showed99%consistent with norB gene of N hamburgensis.Conclusion It was preliminarily confirmed that the microorganism screened in this study might be nitrobac-terium.

Objective To investigate the clinical efficacy of Wumei Wan (Fructus Mume Pills) for diabetic gastroparesis with the syndrome of cold-heat mixture. Methods Sixty-eight diabetic gastroparesis patients with the syndrome of cold-heat mixture were evenly randomized into treatment group and control group. On the basis of treatment for controlling the blood glucose level, the treatment group was given oral use of Wumei Wan and the control group was given oral use of Cisapride. After treatment for one month, the therapeutic effect of both groups was evaluated, and the changes of gastrointestinal symptoms, gastric emptying, 2-hour postprandial glucose(2hPG), glycosylated hemoglobin(HbA1c), and fasting plasma glucose(FPG) in both groups were also observed. Results (1)The total effective rate of the treatment group was 88.24%, and that of the control group was 67.65%, the difference being significant(P<0.05). (2 ) After treatment , the scores of gastrointestinal symptoms of nausea and vomiting, gastric fullness, anorexia, belching and acid regurgitation, abdominal and gastric pain, diarrhea, and constipation in the treatment group were markedly improved(P<0.05 or P<0 . 01 compared with those before treatment ). In the control group, symptom relief was shown in nausea and vomiting, anorexia, and diarrhea(P<0.05). The treatment group had better effect on relieving nausea and vomiting, abdominal and gastric pain, diarrhea, and constipation than the control group(P<0.05 or P<0.01). (3)After treatment, 2hPG, FPG and HbA1c levels were obviously decreased in both groups(P<0.05 compared with those before treatment) , and the decrease in the treatment group was superior to that in the control group (P<0.05). (4)After treatment, half gastric emptying time, full gastric emptying time, and the contraction of gastric antrum were improved in both groups(P<0.05), and the improvement of gastric emptying in the treatment group was superior to that in the cont rol group(P<0.05). Conclusion Wumei Wan has better effect for the treatment of diabetic gastroparesis than western medicine Cisapride.

Objective : To study the protective effects of two antioxidants on the influence of passive smoking on learning and memory ability of mouse offspring. Method: Passive smoking model of pregnant mice was established. Learning and memory ability was evaluated by water maze and long term potentiation (LTP). Nitric oxide (NO) content and nitric oxide synthase (NOS) activity of brain, vitamin E(VE) concentration and reactive oxygen species (ROS) of serum were determined. Results: Latency (the swimming time from beginning to endpoint) and errors (the number of entering blind end) in control and two antioxidant groups were shorter as compared with tobacco smoking (TS) group after 6 d in water maze test, and still shorter after 10 d in control and TS+VE groups. LTP was inhibited in TS group but increased significantly in two antioxidant groups. NOS activitiy was significantly higher in TS group in comparisonwith the control. NO content of TS+VE group was significantly lower than TS and TS+Q groups. Serum VE concentration and ROS level were correlated with the results of latency in water maze and LTP. Conclusion: Passive smoking of the pregnant mice may restrain LTP formation through disturbance of hippocampus function, and reduce the learning and memory ability of the offspring and VE may protect such effects.

New registry classifications of chemical drugs redefined the concepts of new drugs and generic drugs. New drugs emphasize the international innovation including innovate new drugs and modified new drugs. Modified new drugs are improvement of drugs already on the market to achieve superior efficacy. With the advantages of high success rate,high return,low risk and long life cycle,modified new drugs have become the mainstream of new drugs research and development in the world. Modified new drugs R&D in China should be based on clinical demand and focused on the differentiation research in order to obtain new preparations with signif-icant clinical advantages to better serve the clinical practise. This paper states the concept and characteristics of modified new drugs in China,makes case analysis and shares the development history of blockbuster modified new drugs in the world,clarifies the opportuni-ties and challenges and provides ideas for the future development of modified new drugs in China.

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.

AIM:To investigate the effects of down-regulation of protein kinase C (PKC) on the activity of storeoperated Ca2 + channels (SOC) and the proliferation of airway smooth muscle cells (ASMCs). METHODS:Rat bronchial smooth muscle cells were isolated and cultured. Fluo-3 /AM fluorescence was measured by laser confocal microscope to assessing intracellular Ca2 +. Downregulation of PKC activity was achieved by incubation of ASMCs with PKC activator phorbol-12-myristate-13-acetate (PMA,10 ?mol/L) or phorbol 12,13 -dibutyrate (PDBu,1 ?mol/L) for 24 h. The proliferation of ASMCs was assayed by calculating the reduction rates of Alamar blue. RESULTS:Down-regulation of PKC activity by longterm exposure of PMA or PDBu inhibited the proliferation of ASMCs,the similar results were obtained by using PKC inhibitor chelerythrine. Both downregulation of PKC activity and inhibition of PKC activity by chelerythrine reduced Ca2 + entry through SOC channels. Low concentration of PMA (0. 1 ?mol/L) promoted the proliferation of ASMCs,and this effect was inhibited by SOC blocker SKF-96365. CONCLUSION:Inhibition or down -regu-lation of PKC activity results in the inhibition of SOC channels,suggesting that PKC is involved in the activation of these channels. Ca2 + entry through SOC channels might contribute to PKC-promoted proliferation of ASMCs.

Objective To establish a Chinese hamster model of babesia infection, to find the changing pattern of organs and biochemical parameters in Chinese hamster infected with Babesia, and to promote the detection and treatment of babesiosis.Methods Healthy 5-week old Chinese hamsters were infected by intraperitoneal injection of blood containing Babesia.Blood samples were collected at 0, 2, 4, 6, 8, 10, 12, 14, 16, 23, 30, and 37 days after infection from 5 hamsters at each time point.Blood smears were prepared to detect the parasites using Giemsa staining.ELISA assay was employed to test the IL-2 concentration.The blood biochemical indexes were detected using an automatic biochemical analyzer.DNA was extracted from the whole blood and REAL-TIME RCR was performed to determine the reproduction of Babesia.Aftert the animals were sacrificed, the heart, lung, spleen, liver, and kidney were taken to analyze the changes of organ coefficients.Results The highest level of Babesia in the hamsters occurred on day 4 after the Babesia injection, and then showing a decreasing tendency.However, there was a transient increase on the 12th day after infection.The liver and spleen displayed most extensive response to the infection showing hepatomegaly and splenomegaly, but the variation of heart and kidneys coefficients was within the norm.There were prominent changes of blood cells, especially leucocytes, with two peaks at day 10 and 23 after the Babesia infection.The peak changes of blood biochemical indexes occurred at day 12 after infection.The concentration of serum IL-2 reached a peak on the 10th day after infection.Conclusions The Chinese hamsters display typical characteristics of tick-borne diseases such as hepatomegaly and splenomegaly.The immunological system is activated along with the infection and reaches a highest stage in the second week.Afterwards the Babesia can live in the hamster body for a long period of time.The results of this study provide useful information supporting further studies on the detection, treatment and prevention of Babesiosis.

Objective To summarize the clinical characteristics,diagnosis and treatment of malignant tumor after renal transplantation.Methods The clinical data of 2106 renal transplants in 1945 patients undertaken in our hospital from September 1978 to December 2009 was retrospectively studied.Results Of these 1945 patients, 39 cases were diagnosed as having malignant tumor (incidence: 2.0 ％).The interval between transplantation and clinical diagnosis ranged from 8 to 124 months with a median of 57.0 months.Among the 39 cases of malignancy, there were 22 urinary system carcinomas, 8 digestive system carcinomas,2 lung cancers,2 breast cancers,2 lymphomas,1 dura small cell carcinoma,1 pleura poorly differentiated carcinoma and 1 metastatic carcinoma of liver with unknown primary tumor. Surgery was conducted in 28 patients,of which 16 were survived but the other 12 patients died of metastasis ranged from 3 months to 96 months (median,33 months) after operation.11 cases without operation died within from 3 d to 36 months (median,5 months) after diagnosis.Conclusions The incidence of malignant tumors in renal transplant recipients increased markedly.The most common type of the malignant tumors is urinary system carcinoma.The key measure of success in treating malignancy after renal transplantation is early diagnosis and surgical resection.

Objective To explore the early-term efficacy of porous titanium trabecular metal acetabular (TTM) compo-nents produced by E-beam technology in total hip arthroplasty(THA). Methods From May 2012 to JUN 2013, nineteen primary THA patients (20 hips) were devided into observation group of porous TTM acetabular prosthesis produced by E-beam technology and control group of Pinnacle acetabular prosthesis. TTM group had nine patients (10 hips), including 7 males and 2 females, with the mean age of 61.0 years. Pinnacle group had ten patients (10 hips), including 6 males and 4 females, with the mean age of 58.6 years. The patients were followed up at the 6,12,24 weeks after operation. X-rays were taken at the follow-ups. Acetabular compo-nent stability and osseointegration were assessed through radiograph. Harris scores system,SF-36 Heath Survey Questionnaire and WOMAC Osteoarthritis index were recorded before and after operation to evaluate clinical effects of surgery. Results In TTM group, the Harris score improved from 48.2 ± 5.5 preoperatively to 89.5 ± 4.0 postoperatively, SF-36 scores from 329.1 ± 86.7 to 763.8 ± 15.1, and WOMAC index from 18.8 ± 11.0 to 1.3 ± 0.9. No cases suffered aseptic loosening and migration. After 24 weeks, There was no significant difference regarding Harris scores system,SF-36 Heath Survey Questionnaire and WOMAC Os-teoarthritis index between two groups. In TTM group, two hips had a radiolucent line postoperatively, which occurred in 2 and 3 zone. The lines were disappeared 24 weeks later. Conclusion In THA, the initial stability of TTM produced by E-beam tech-nology was fine, which can facilitate bone ingrowth in early stage. However, the long-term follow-up effect still needs to be con-firmed.